Detection of chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes.
We analyzed the distribution of chromogranins A and B in normal and neoplastic endocrine tissues with secretory granules using 35S-labeled and biotin-labeled oligonucleotide probes by in situ hybridization (ISH). Both radioactive and nonradioactive probes detected messenger RNAs (mRNAs) in frozen and paraffin tissue sections. Endocrine tissues with variable immunoreactivities for chromogranin A protein, such as small-cell lung carcinomas, neuroblastomas, insulinomas, and parathyroid adenomas, expressed the mRNA for chromogranins A and B in most cells. Some technical problems with the biotinylated probes included nonspecific nuclear staining and endogenous alkaline phosphatase, which was not completely abolished by levamisole pretreatment. A differential distribution of chromogranins A and B was seen in pituitary prolactinomas, which expressed abundant chromogranin B but not chromogranin A mRNAs, and in parathyroid adenomas, which expressed abundant chromogranin A but only small amounts of chromogranin B mRNAs. These results indicate that ISH can be used to detect chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes and that the mRNAs for chromogranin A and B are demonstrable in some tumors even when the chromogranin proteins cannot be detected by immunohistochemistry.
Lloyd
R V
RV
Department of Pathology, University of Michigan, Ann Arbor 48109-0054.
Jin
L
L
Fields
K
K
D001710:Biotin / Q000032:analysis*; D015152:Blotting, Northern; D002864:Chromogranins / Q000032:analysis*; D004702:Endocrine Glands / Q000032:analysis*; D004700:Endocrine System Diseases / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000032:analysis*; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000032:analysis; D012334:RNA, Neoplasm / Q000032:analysis; D013462:Sulfur Radioisotopes
D002864:Chromogranins; D009419:Nerve Tissue Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D013462:Sulfur Radioisotopes; D001710:Biotin